The present invention relates to a method for analyzing a metabolic flux, that is, a metabolic flux analysis method, a program for the method and a recording medium recording the program. Specifically, the present invention relates to a metabolic flux analysis method using an isotope-labeled substance, a program for the method and a recording medium recording the program.
The metabolic flux analysis method is a method for quantitatively determining an intracellular metabolic flux by analyzing intracellular balances of metabolites or isotope-labeled compounds and conducting isotope compound tracer experiments with an analytical technique such as nuclear magnetic resonance (NMR) or mass spectrometry (MS). In recent years, this method has drawn attentions as a technique for stoichiometrically analyzing the quantitative ratio of metabolites (carbon balance) in metabolic pathways in an objective cell (Metabolic Engineering 3, 265-283, 2001).
Various studies are being conducted to develop an accurate analytical technique for use in metabolic flux analyses. The theory concerning metabolic flux analysis using isotope-labeled substrates has been reported in many papers and is being established (Biotechnology and Bioengineering 55, 101-117, 1997. Biotechnology and Bioengineering 55, 118-13, 1997% Biotechnology and Bioengineering 66, 69-85, 1999% Biotechnology and Bioengineering 66, 86-103, 1999). Although many experiments are being conducted to establish a metabolic flux analysis method, researches based on a continuous culture method utilizing a synthetic medium as an ideal condition are common to obtain high analytical precision (Journal of Biological Chemistry 275, 35932-35941, 2000). Further, although there are a few reports on metabolic flux analysis performed by batch culture as a more practical culture method, only isotope distributions of several substances discharged in a medium have been measured, and no calculation has been performed at all based on the measurement of isotope distributions in intracellular substances (European Journal of Biochemistry 268, 2441-2455, 2001). Isotope substrates are generally expensive and when an experiment is conducted according to the conventional method, the considerably increasing costs of the experiment become a problem (Journal of Biotechnology 94, 37-63, 2002% Metabolic Engineering 3, 195-205, 2001).